Microwave Devices for Biosensors and Single Cell Dielectric Spectroscopy

用于生物传感器和单细胞介电谱的微波设备

• 批准号: RGPIN-2019-05859
• 负责人: Bridges, Greg
• 金额: $ 3.35万
• 依托单位: University of Manitoba
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=738551
• 关键词: Microwave; Devices; Biosensors; Single; Cell

Studying the physiology of biological cells and monitoring their response to different stimuli or drugs is essential in bioprocess and biomedical research. Many currently used instruments for cellular analysis utilize chemical markers, sophisticated equipment and time-consuming preparation techniques. Microfluidic devices employing dielectric analysis are a promising alternative as they are label-free, can be miniaturized and use nanoliter fluid volumes. The objectives of the proposed research program is to develop microfluidics devices integrated with microwave dielectric sensors and actuators for in-flow single cell analysis, use these to study cell response to different types of stress, develop new cell dielectric models, and relate cell dielectric properties to physiology. Tissues, cells and other biomedia have unique dielectric properties that can be used to provide a wealth of information, such as a cell's growth phase, whether it is healthy or cancerous, or if drug-induced changes have occurred. Most current methods for measuring single cell dielectric properties employ impedance or dielectrophoresis at low frequencies using only one or two frequencies. Much additional information can be gained by measuring the complete spectrum and extending this to microwave frequencies, a regime where fields can penetrate the cell interior and the polarizability of large molecules can be detected. Dielectric spectroscopy of single cells is extremely difficult due to their small size. We will explore new microwave dielectrophoresis sensing techniques to measure single cells over a wide frequency spectrum and use this to develop multi-parameter cell dielectric models. Microfliudic devices will be used to study cell response to starvation, heat shock and microwave electroporation (EP). Detection of starvation-induced apoptosis is important in biopharmaceutical bioprocessing and biomedical research. Dielectric changes to cells under starvation will be related to cell physiology and the developed cell models will be used to understand bulk-suspension dielectric measurements. Heat shock and exposure to high-intensity electric fields (inducing EP) can permanently or temporarily permeabilize cell membranes. This is employed for transfection, enhancing drug uptake in cancer therapy, inducing apoptosis, or in the extreme case, lysis. Microfluidic devices will be developed to apply and simultaneously measure the dielectric changes to cells associated with these two stressors. Of particular interest is CW microwave EP, where we will explore use of low-intensity fields to induce EP effects. The outcomes of the research will be new microfluidic single cell dielectric spectroscopy tools integrated with stimulus capabilities, providing new non-invasive label-free single cell analysis instruments. The research program will provide both theoretical and experimental training to highly qualified personnel in an interdisciplinary environment.

研究生物细胞的生理学并监测它们对不同刺激或药物的反应对于生物过程和生物医学研究至关重要。目前许多用于细胞分析的仪器利用化学标记、精密设备和耗时的制备技术。采用介电分析的微流体装置是一种有前途的替代方案，因为它们无需标记，可以小型化并使用纳升流体体积。拟议研究计划的目标是开发与微波介电传感器和执行器集成的微流体器件，用于流动单细胞分析，利用这些器件来研究细胞对不同类型应力的响应，开发新的细胞介电模型，并关联细胞介电特性到生理学。组织、细胞和其他生物介质具有独特的介电特性，可用于提供丰富的信息，例如细胞的生长期、健康还是癌变，或者是否发生了药物引起的变化。当前测量单细胞介电特性的大多数方法仅使用一种或两种频率在低频下采用阻抗或介电泳。通过测量完整的光谱并将其扩展到微波频率，可以获得许多额外的信息，在这种情况下，场可以穿透细胞内部，并且可以检测大分子的极化率。由于单细胞尺寸小，介电谱分析极其困难。我们将探索新的微波介电泳传感技术来测量宽频谱上的单细胞，并用它来开发多参数细胞介电模型。 微流体装置将用于研究细胞对饥饿、热休克和微波电穿孔（EP）的反应。检测饥饿诱导的细胞凋亡对于生物制药生物加工和生物医学研究非常重要。饥饿条件下细胞的介电变化将与细胞生理学相关，开发的细胞模型将用于理解体悬浮介电测量。热休克和暴露于高强度电场（诱导 EP）可以永久或暂时地使细胞膜通透。它用于转染、增强癌症治疗中的药物摄取、诱导细胞凋亡，或者在极端情况下用于裂解。将开发微流体装置来应用并同时测量与这两种应激源相关的细胞的介电变化。特别令人感兴趣的是 CW 微波 EP，我们将探索使用低强度场来诱导 EP 效应。该研究的成果将是集成了刺激功能的新型微流控单细胞介电光谱工具，提供新型非侵入性无标记单细胞分析仪器。该研究计划将在跨学科环境中为高素质人才提供理论和实验培训。